Contextual Modulation of Social and Endocrine Correlates of Fitness: Insights From the Life History of a Sex Changing Fish

Steroid hormones are critical regulators of reproductive life history, and the steroid sensitive traits (morphology, behavior, physiology) associated with particular life history stages can have substantial fitness consequences for an organism. Hormones, behavior and fitness are reciprocally associated and can be used in an integrative fashion to understand how the environment impacts organismal function. To address the fitness component, we highlight the importance of using reliable proxies of reproductive success when studying proximate regulation of reproductive phenotypes. To understand the mechanisms by which the endocrine system regulates phenotype, we discuss the use of particular endocrine proxies and the need for appropriate functional interpretation of each. Lastly, in any experimental paradigm, the responses of animals vary based on the subtle differences in environmental and social context and this must also be considered. We explore these different levels of analyses by focusing on the fascinating life history transitions exhibited by the bi-directionally hermaphroditic fish, Lythrypnus dalli. Sex changing fish are excellent models for providing a deeper understanding of the fitness consequences associated with behavioral and endocrine variation. We close by proposing that local regulation of steroids is one potential mechanism that allows for the expression of novel phenotypes that can be characteristic of specific life history stages. A comparative species approach will facilitate progress in understanding the diversity of mechanisms underlying the contextual regulation of phenotypes and their associated fitness correlates

Rapid social regulation of 3β-HSD activity in the songbird brain

Rapid increases in plasma androgens are generally associated with short-term aggressive challenges in many breeding vertebrates. However, some animals such as song sparrows (Melospiza melodia) are aggressive year-round, even during the non-breeding season, when gonads are regressed and systemic testosterone (T) levels are non-detectable. In contrast, levels of the prohormone dehydroepiandrosterone (DHEA) are elevated year-round in the plasma and brain. The local conversion of brain DHEA to potent androgens may be critical in regulating non-breeding aggression. 3β-hydroxysteroid dehydrogenase/Δ4-Δ5 isomerase (3β-HSD) catalyzes DHEA conversion to androstenedione (AE) and the cofactor NAD⁺ assists in this transformation. In this thesis, I asked whether brain 3β-HSD activity is regulated by social encounters in seasonally breeding male songbirds. In Experiment 1, I looked at the long-term seasonal regulation of brain 3β-HSD activity. 3β-HSD activity was highest in the non-breeding season compared to the breeding season and molt. In Experiment 2, I hypothesized that brain 3β-HSD activity is rapidly regulated by short-term social encounters during the non-breeding season. A 30 min social challenge increased aggressive behavior. Without exogenous NAD⁺, there was ~355% increase in 3β-HSD activity in the caudal telencephalon and ~615% increase in the medial central telencephalon compared to controls (p<0.05). With exogenous NAD⁺, there was no effect of social challenge on 3β-HSD activity. These data suggest that endogenous cofactors play a critical role in the neuroendocrine response to social challenges. The increase in brain DHEA conversion to AE during social challenges may be a mechanism to rapidly increase local androgens in the non-breeding season, when there are many costs of systemic T.Science, Faculty ofZoology, Department ofGraduat

Contextual modulation of social and endocrine correlates of fitness: insights from the life history of a sex changing fish

Steroid hormones are critical regulators of reproductive life history, and the steroid sensitive traits (morphology, behavior, physiology) associated with particular life history stages can have substantial fitness consequences for an organism. Hormones, behavior and fitness are reciprocally associated and can be used in an integrative fashion to understand how the environment impacts organismal function. To address the fitness component, we highlight the importance of using reliable proxies of reproductive success, when studying proximate regulation of reproductive phenotypes. To understand the mechanisms by which the endocrine system regulates phenotype, we discuss the use of particular endocrine proxies and the need for appropriate functional interpretation of each. Lastly, in any experimental paradigm, the responses of animals vary based on the subtle differences in environmental and social context and this must also be considered. We explore these different levels of analyses by focusing on the fascinating life history transitions exhibited by the bi-directionally hermaphroditic fish, Lythrypnus dalli. Sex changing fish are excellent models for providing a deeper understanding of the fitness consequences associated with the behavioral and endocrine variation. We close by proposing that local regulation of steroids is one potential mechanism that allows for the expression of novel phenotypes that can be characteristic of specific life history stages. A comparative species approach will facilitate progress in understanding the diversity of mechanisms underlying the contextual regulation of phenotypes and their associated fitness correlates

The Plasticity of Social Status: Stress Hormones in a Hermaphroditic Fish

Psychosocial stress due to dominance rank can have profound effects on physiology, behavior, and metabolism. Cortisol, a hormone that controls stress-induced responses in all vertebrates, may be an important mediator of changes in behavioral phenotype due to social structure or changes in status, such as dominance and/or subordinate rank. Bluebanded gobies, Lythrypnus dalli, are bidirectionally hermaphroditic fish, in which sex change occurs due to a change in social structure. In stable groups, L. dalli live in linear social dominance hierarchies, such that one male dominates over many females and defends a nesting territory. Upon male removal (MR), the most dominant female exhibits rapid increases in rates of aggressive and territorial behavior. This species is ideal for exploring the effect of social status and instability on cortisol levels due to the plasticity of life history transitions, such as sex change. Here, we compared systemic cortisol levels amongst the group hierarchy in stable groups, 30 min after MR, and 24 h after MR. Behavioral observations were taken in 10 min bins and used to determine the status of each fish in a group. There was no significant difference in water-borne cortisol level in stable groups or after male removal. These data indicate that systemic cortisol levels are not associated with psychosocial stress. Future studies will investigate more specific measures from brain, such as cortisol, cortisol producing enzymes, or glucocorticoid receptor levels

11β-HSD Types 1 and 2 in the Songbird Brain

Glucocorticoid (GC) hormones act on the brain to regulate diverse functions, from behavior and homeostasis to the activity of the hypothalamic–pituitary–adrenal axis. Local regeneration and metabolism of GCs can occur in target tissues through the actions of the 11β-hydroxysteroid dehydrogenases [11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1) and 11 beta-hydroxysteroid dehydrogenase type 2 (11β-HSD2), respectively] to regulate access to GC receptors. Songbirds have become especially important model organisms for studies of stress hormone action; however, there has been little focus on neural GC metabolism. Therefore, we tested the hypothesis that 11β-HSD1 and 11β-HSD2 are expressed in GC-sensitive regions of the songbird brain. Localization of 11β-HSD expression in these regions could provide precise temporal and spatial control over GC actions. We quantified GC sensitivity in zebra finch (Taeniopygia guttata) brain by measuring glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) expression across six regions, followed by quantification of 11β-HSD1 and 11β-HSD2 expression. We detected GR, MR, and 11β-HSD2 mRNA expression throughout the adult brain. Whereas 11β-HSD1 expression was undetectable in the adult brain, we detected low levels of expression in the brain of developing finches. Across several adult brain regions, expression of 11β-HSD2 covaried with GR and MR, with the exception of the cerebellum and hippocampus. It is possible that receptors in these latter two regions require direct access to systemic GC levels. Overall, these results suggest that 11β-HSD2 expression protects the adult songbird brain by rapid metabolism of GCs in a context and region-specific manner

Data from: Agonistic reciprocity is associated with reduced male reproductive success within haremic social networks

While individual variation in social behaviour is ubiquitous and causes social groups to differ in structure, how these structural differences affect fitness remains largely unknown. We used social network analysis of replicate bluebanded goby (Lythrypnus dalli) harems to identify the reproductive correlates of social network structure. In stable groups, we quantified agonistic behaviour, reproduction and steroid hormones, which can both affect and respond to social/reproductive cues. We identified distinct, optimal social structures associated with different reproductive measures. Male hatching success (HS) was negatively associated with agonistic reciprocity, a network structure that describes whether subordinates ‘reciprocated’ agonism received from dominants. Egg laying was associated with the individual network positions of the male and dominant female. Thus, males face a trade-off between promoting structures that facilitate egg laying versus HS. Whether this reproductive conflict is avoidable remains to be determined. We also identified different social and/or reproductive roles for 11-ketotestosterone, 17β-oestradiol and cortisol, suggesting that specific neuroendocrine mechanisms may underlie connections between network structure and fitness. This is one of the first investigations of the reproductive and neuroendocrine correlates of social behaviour and network structure in replicate, naturalistic social groups and supports network structure as an important target for natural selection

11β-HSD Types 1 and 2 in the Songbird Brain

Glucocorticoid (GC) hormones act on the brain to regulate diverse functions, from behavior and homeostasis to the activity of the hypothalamic-pituitary-adrenal axis. Local regeneration and metabolism of GCs can occur in target tissues through the actions of the 11β-hydroxysteroid dehydrogenases [11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1) and 11 beta-hydroxysteroid dehydrogenase type 2 (11β-HSD2), respectively] to regulate access to GC receptors. Songbirds have become especially important model organisms for studies of stress hormone action; however, there has been little focus on neural GC metabolism. Therefore, we tested the hypothesis that 11β-HSD1 and 11β-HSD2 are expressed in GC-sensitive regions of the songbird brain. Localization of 11β-HSD expression in these regions could provide precise temporal and spatial control over GC actions. We quantified GC sensitivity in zebra finch (Taeniopygia guttata) brain by measuring glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) expression across six regions, followed by quantification of 11β-HSD1 and 11β-HSD2 expression. We detected GR, MR, and 11β-HSD2 mRNA expression throughout the adult brain. Whereas 11β-HSD1 expression was undetectable in the adult brain, we detected low levels of expression in the brain of developing finches. Across several adult brain regions, expression of 11β-HSD2 covaried with GR and MR, with the exception of the cerebellum and hippocampus. It is possible that receptors in these latter two regions require direct access to systemic GC levels. Overall, these results suggest that 11β-HSD2 expression protects the adult songbird brain by rapid metabolism of GCs in a context and region-specific manner

Anti-Müllerian Hormone Levels in the Gonads of Sex-Changing Fish

Anti-Müllerian Hormone Levels in the Gonads of Sex-Changing Fish Anti-Müllerian Hormone (AMH) promotes the development of the testes through the suppression of the female ovaries in the developing embryo. This glycoprotein has also been found to be a key contributor to follicular development in adult females of different species including humans. The purpose of this study was to determine the presence of AMH in a hermaphroditic fish, the blue-banded goby, (Lythrypnus dalli). While most species only have AMH present in embryos and adult females, L. dalli and other sex-changing fish have AMH in adult males as well. For this study two levels of analyses were used 1) Ovarian RNA was extracted and converted into cDNA which will be used to investigate AMH and AMH receptor mRNA levels in future studies; 2) Ovarian tissue of adult females, testes of adult males, and bipotential gonads of juveniles were homogenized, purified through solid-phase extraction and analyzed using a specific AMH ELISA. We found that there was almost three times as much AMH in males than in females, and AMH in females was roughly half of the AMH found in juveniles. Ongoing studies will explore the role of AMH during sexual differentiation of juveniles and adult sex change

Stuck in a Bucket: The Effect of Confinement Stress on Cortisol Levels in Brook Trout (\u3cem\u3eSalvelinus fontinalis\u3c/em\u3e)

In vertebrates, a primary physiological stress response is characterized by a rapid increase in circulating glucocorticoid levels. Repeated exposure to external stressors can have an impact on overall health, such as metabolism, growth, and immunity. Salmonids are a cold-water fish found in higher elevations; they are important both environmentally and recreationally. Brook Trout, Salvelinus fontinalis, prefer open, clean environments but are often exposed to stressful confinement in hatcheries. The goal of this study is to examine the primary stress response that fish experience during confinement. We investigated the effect of physical and visual stressors on cortisol levels in fingerling brook trout. Wild-caught fish were acclimated in a lab for 6 months. On the day of the experiment, we compared two different routes by which systemic cortisol levels could be collected: water-borne and plasma. On the day of the experiment, fish were held in a smaller confined bucket for one of the following time periods: 3, 5, 15, 30, or 60 mins and then a blood sample was obtained. Steroid samples will be extracted from both water and plasma through solid phase extraction using C18 columns and a vacuum manifold, and measured using an enzymeimmunoassay. We predict a steady increase in cortisol levels over the five-time points. These data will be important to determine the time frame of the stress response resulting from confinement stress and help assess other negative effects of chronic stress in salmonids

3β-HSD expression in the CNS of a manakin and finch

The prohormone, dehydroepiandrosterone (DHEA) circulates in vertebrate blood with the potential for actions on target tissues including the central nervous system (CNS). Many actions of DHEA require its conversion into more active products, some of which are catalyzed by the enzyme 3β-hydroxysteroid-dehydrogenase/isomerase (3β-HSD). Studies of birds show both expression and activity of 3β-HSD in brain and its importance in regulating social behavior. In oscine songbirds, 3β-HSD is expressed at reasonably high levels in brain, possibly linked to their complex neural circuitry controlling song. Studies also indicate that circulating DHEA may serve as the substrate for neural 3β-HSD to produce active steroids that activate behavior during non-breeding seasons. In the golden-collared manakin (Manacus vitellinus), a sub-oscine bird, low levels of courtship behavior are displayed by males when circulating testosterone levels are basal. Therefore, we asked whether DHEA circulates in blood of manakins and whether the brain expresses 3β-HSD mRNA. Given that the spinal cord is a target of androgens and likely important in regulating acrobatic movements, we also examined expression of this enzyme in the manakin spinal cord. For comparison, we examined expression levels with those of an oscine songbird, the zebra finch (Taeniopygia guttata), a species in which brain, but not spinal cord, 3β-HSD has been well studied. DHEA was detected in manakin blood at levels similar to that seen in other species. As described previously, 3β-HSD was expressed in all zebra finch brain regions examined. By contrast, expression of 3β-HSD was only detected in the manakin hypothalamus where levels were greater than zebra finches. In spinal cord, 3β-HSD was detected in some but not all regions in both species. These data point to species differences and indicate that manakins have the substrate and neural machinery to convert circulating DHEA into potentially active androgens and/or estrogens